Relating to organic compounds

ABSTRACT

A cosmetic ingredient comprising crenatoside and acteoside when applied to the skin is useful to provide cosmetic and perfumery benefits.

The present invention is concerned with cosmetic ingredients, which maybe applied topically to the skin in order to provide beneficial effectsrelated to perfumery or to the preservation of skin beauty over time.The invention is particularly concerned with said cosmetic ingredientsthat are able to exert beneficial effects by influencing the compositionof the skin microbiota.

In our 20s, our skin renews itself approximately once every 27 days,whereas in our 50s, this same process takes up to 39 days. In otherwords, in only a matter of a few decades, our skin needs about 44% moretime to renew itself.

The skin regeneration process is complex. The process of maintainingskin health and beauty begins with stem cell activation in the basallayer of the skin, upwards through to the superficial layers of thestratum corneum and the stratum microbium.

The stratum microbium is a term that refers to the massive amount ofbeneficial microorganisms that colonize the surface of our skin, andalthough it is not technically a layer of skin, it does interactsymbiotically with the skin to enable individuals to maintain theirhealth on a daily basis. The stratum microbium remains in a state offlux, in which the composition of the resident microflora (ormicrobiota) changes over time in response to factors both intrinsic andextrinsic to the host. It is now recognized that a healthy, balancedmicrobiota can provide not only health benefits, but also cosmeticbenefits to the human host.

There is a need to improve the health and/or appearance of human skin byproviding cosmetic ingredients as well as methods of cosmeticintervention that can provide beneficial effects, such as acceleratingthe skin rejuvenation process, and can deliver targeted biologicalaction holistically from the lower to the higher layers of theepidermis, as well as promote a healthy, balanced microbiota.

The applicant has surprisingly discovered a cosmetic ingredient thataddresses the deficiencies of the prior art.

Accordingly, the invention provides in a first aspect a cosmeticingredient comprising a mixture of crenatoside and acteoside.

In another aspect, the invention provides a method of improving thecondition or appearance of human skin, the method comprises the step oftopically applying to the skin the cosmetic ingredient.

In another aspect, the invention provides a method of balancing the skinmicrobiota of a human host, the method comprising the step of topicallyapplying to the skin the cosmetic ingredient.

In another aspect, the invention provides a formulation suitable fortopical application to the skin containing the cosmetic ingredient.

These and other aspects of the invention will be further understood inview of the following detailed description of particular embodiments ofthe invention.

The cosmetic ingredient of the present invention may be a plant extractenriched in crenatoside and acteoside.

The plant extract may be obtained from plants of the Orobanchaceaefamily, and more particularly from Orobanche rapum.

Although the compounds crenatoside and acteoside are both water-soluble,the applicant found that they were unstable in water. Consequently, theextract is preferably obtained by extracting the compounds from plantmaterial using an alcohol, and more particularly ethanol. It isparticularly preferred if the alcohol is water-free.

In order to ensure that the extract is stable in terms of both actives'content and colouration, it is desirable that the cosmetic ingredient ofthe present invention is presented as an extract in a hydric solvent.

A hydric solvent, as the term is used in the present invention, refersto a solvent that contains at least two hydroxyl groups. Suitable hydricsolvents include glycerol or glycols, such as dipropylene glycol,diethylene glycol, dibutylene glycol, propylene glycol, butylene glycol,pentylene glycol, heptylene glycol, and hexylene glycol.

A particularly preferred solvent for use in the extract is1,3-propanediol.

The plant material can be extracted directly using a hydric solventreferred to above. However, it is preferred if an alcohol, such asethanol, is employed as the extracting medium, and a subsequentsolvent-exchange step is carried out to replace the extraction solventwith a hydric solvent in order to stabilize the cosmetic ingredient. Theextraction solvent can be removed in a relatively straightforward mannerif it is a lower boiling solvent compared to the hydric solvent.

In an embodiment of the present invention, the cosmetic ingredient inthe form of a plant extract may be obtained in a process comprising thesteps of:

-   -   I) Extracting a plant source containing the compounds        crenatoside and acteoside in an extraction solvent, preferably        ethanol;    -   II) Filtering the extract to remove solids;    -   III) Carrying out a solvent-exchange step comprising mixing the        extract with a hydric solvent and removing the extraction        solvent by evaporation;    -   IV) Optionally decolourizing the extract obtained in step III),        using a suitable decolourizing agent, such as activated        charcoal;    -   V) Filtering the extract to remove solids; and    -   VI) Optionally adjusting the extract to the desired dry matter        content by evaporation.

The cosmetic ingredient of the present invention is characterized by avery high content of crenatoside and acteoside. In an embodiment of theinvention, the cosmetic ingredient in the form of a plant extractcontains at least 15 wt %, more particularly at least 16 wt %, moreparticularly at least 17 wt %, more particularly at least 18 wt %, andmore particularly at least 19 wt % of a mixture of crenatoside andacteoside, based on the total weight of the dry matter of the extract.In an embodiment, said mixture comprises about 20 wt % to about 60 wt %of crenatoside and about 80 wt % to about 40 wt % of acteoside, based onthe total weight of said mixture, preferably about 30 wt % to about 50wt % of crenatoside and about 70 wt % to about 50 wt % of acteoside,more preferably about 35 wt % to about 45 wt % of crenatoside and about65 wt % to about 55 wt % of acteoside, for example about 40 wt % ofcrenatoside and about 60 wt % of acteoside. It is also possible that thecosmetic ingredient of the present invention contains at least 15 wt %,more particularly at least 16 wt %, more particularly at least 17 wt %,more particularly at least 18 wt %, and more particularly at least 19 wt% of each of crenatoside and acteoside, based on the total weight of thedry matter of the extract.

It is an advantage of the present invention that the cosmetic ingredientis easily incorporated into all manner of formulations suitable for usein topical applications, including cosmetic preparations and finefragrances.

Since highly coloured ingredients may be unacceptable as raw materialsfor manufacturers of cosmetic preparations or fine fragrances, it ishighly advantageous that the cosmetic ingredient according to theinvention can be prepared such that its colour or hue will not alter orimpair the colour tone of any cosmetic preparation or fine fragranceinto which they might be incorporated at levels required to deliver thedesired cosmetic or other beneficial effect.

It is a particular advantage of the present invention that cosmeticingredients in the form of plant extracts can be obtained having arequisite lightness of colour or hue, which enables them to be used atefficacious levels in cosmetic preparations of fine fragranceapplications, without affecting the tone of the cosmetic preparation orfine fragrance.

The lightness and hue of the cosmetic ingredient can be characterized byCIELAB chromaticity coordinates L* a* b* according to colorimetricmethods known in the art. The L* value is a value specifying thelightness of a substance and is indicated by a value between 0 and 100.An L* value of 100 indicates the brightest state (completely white), andan L* value of 0 indicates the darkest state (completely black). The b*value specifies the blue-yellow hue of a substance. The larger is the b*value, the higher is the degree of yellowness. The smaller is the b*value, the higher is the degree of blueness.

The L* a* b* values of a sample may be measured using any suitablecommercially available spectrophotometer, such as a Lico 690. Thespectrophotometer is typically powered up for at least one hour beforemaking a measurement. A glass container provided therefor should be halffilled with a sample to be measured taking care to ensure that thebottom of the container is fully covered by the sample. Thereafter, thefilled container should be placed in the sample stand provided therefor.The sample key on the instrument should be pressed and the L* a* and b*values read off the display panel. Before taking any readings, theinstrument should be calibrated for the zero and 100% reflection byplacing black and white objects provided therefor on the window of theinstruments optical sensor.

In an embodiment of the invention, the cosmetic ingredient has at leastone of an L* value of at least about 70; an a* value greater than about4 and less than about 19; and a b* value greater than about 78 and lessthan about 91, wherein L*, a* and b* values represent the CIELABsystem's chromaticity coordinates.

The cosmetic ingredient according to the present invention can exert areactivation effect on the natural skin renewal cycle from top tobottom, that is from the superficial layers of the stratum corneum andthe stratum microbium to the stem cells in the basal layer of the skin.

In particular, the applicant has demonstrated that the cosmeticingredient is multi-functional in that it can exert at least one,preferably all, of the following holistic effects on the natural skinrenewal cycle:

-   -   The cosmetic ingredient of the present invention can provide        protection to epidermis stem cells.    -   The cosmetic ingredient of the present invention can deliver        anti-apoptosis activity.    -   The cosmetic ingredient of the present invention can stimulate        epidermis metabolism.    -   The cosmetic ingredient of the present invention can stimulate        epidermis cell differentiation.    -   The cosmetic ingredient of the present invention can stimulate        the production of the skin's barrier components.    -   The cosmetic ingredient of the present invention can activate        the process of natural exfoliation.    -   The cosmetic ingredient of the present invention can act to        maintain the balance of the skin microflora (microbiota).

More particularly, the cosmetic ingredient of the present invention canhelp stem cells maintain their capability to replicate over time. Thiseffect was demonstrated by observing the clone-forming capacity of stemcells over time when cultivated with and without the cosmetic ingredientusing optical microscopy. Still further, in vitro studies examining theeffect on human stem cells of daily UV exposure with or without thepresence of the cosmetic ingredient demonstrated that levels of theapoptosis inhibiting protein survivin were significantly higher insamples exposed to the cosmetic ingredient.

More particularly, the cosmetic ingredient of the present invention hasbeen shown in studies to inhibit the enzymes Caspase 9 and Caspase 3,both of which are pro-apoptotic enzymes.

More particularly, studies on human skin explants from healthy donorsuntreated and treated with the cosmetic ingredient of the presentinvention demonstrate that samples treated with the cosmetic ingredientshow increased expression of Ki67 positive cells (measured byimmunohistochemistry), which indicates increased cellular metabolism inthe epidermis.

More particularly, transcriptomic analysis of epidermis differentiationgenes demonstrated stimulation of the expression of certain proteinsassociated with the epidermis cell differentiation. These proteinsincluded filaggrin, involucrin and loricrin.

More particularly, the cosmetic ingredient of the present invention canincrease ceramide production, demonstrating the capability to restorethe barrier function of the skin.

More particularly, the cosmetic ingredient of the present invention canstimulate the production of the enzyme Kallikrein 5 protease, which isinvolved in the desquamation process.

The surface of human skin is home to a wide variety of microorganisms.Collectively, these microorganisms form a microbiome, often referred toas the skin's microbiota. It is generally believed that skin health andappearance requires a balanced collection of skin microorganisms.However, it is known that the balance of the skin's microflora canchange over time for intrinsic and extrinsic reasons. For example,certain undesirable microorganisms, such as pathogenic bacteria, moldsand yeasts, may attempt to colonize the skin to the detriment of theresident microorganisms; or nutrients that reach the surface of the skinmay be effective for some microorganisms, but not others; or sanitizingproducts that a human host applies to its skin may be relatively moredeleterious for some microorganisms than others.

The applicant found, however, that the balance of skin microfloraevolved differently over time depending upon whether the skin surfacewas treated with the cosmetic ingredient of the present invention, orremained untreated. More particularly, the applicant undertook ametagenomic study by swabbing, extracting and sequencing the totalmicrobial DNA of healthy volunteers (women, forearm skin) at a timeT=zero and after 14 days, and found that the cosmetic ingredientsubstantially balanced the compositional make-up of the microbiota,which contained as the main phyla, in terms of relative abundance,Actinobacteria, Cyanobacteria, Proteobacteria, Chloroflexi, Firmicutes,Saccharibacteria, and Bacteroidetes.

By balancing the skin's microbiota favourably in this way, the cosmeticingredient of the present invention offers beneficial effects for thehost, including improvements in skin health and appearance, as well aspotentially alleviating a number of skin disorders.

As used herein, the term “balance” as it relates to the microbiota meansthat the difference in the amount of at least one of the main phylareferred to above on skin treated with the cosmetic ingredient over anappropriate time interval, e.g. 2 weeks, does not change as much (eitherpositively or negatively) as compared to an untreated control. Themicrobiota is also considered balanced if, during the same timeinterval, the growth of a pathogen (e.g. Finegoldia magna) is less onskin treated with the cosmetic ingredient as compared to an untreatedcontrol.

A method of assessing the balance of the microbiota is set forth in moredetail in the examples hereinbelow.

A balanced microbiota can support the health and condition of human skinby means of a number of passive and active mechanisms: Residentmicroorganisms may passively compete with pathogens for niches on theskin surface or consume nutrients essential for the growth of pathogens.They also actively consume material littering the skin, comprising amixture of bodily excretions as well as material that is applied to theskin or comes into contact with the skin from external sources, toproduce a complex mixture of metabolites and by-products that caninhibit or even kill pathogens.

As a result of the interaction between the microbiota and this cutaneousmaterial, the surface of the skin is home to a mixture of chemicalcompounds including proteins, lipids, and carbohydrates, as well asmetabolites and by-products, which may comprise alcohols, fatty acids,aldehydes, and the like. This complex chemical environment may containcompounds that are volatile and odorous, and which can contribute to thecharacteristic odour, or malodour, of individuals. Still further, thissurface chemistry can influence the manner in which the headspace of aperfume composition applied to the skin will evolve over a period oftime.

Perfume ingredients are oils, and the tendency with which they willeither rapidly diffuse into the headspace above the skin or partitioninto the superficial layers of the skin and evaporate slowly will dependto some extent upon the composition of the skin surface chemistry, whichis directly influenced by the balance of the microbiota. For example,the hydrophobic/hydrophilic balance of the chemical mixture may causesome perfume ingredients to bloom rapidly into the headspace, and othersto become fixed to the surface. Other perfume ingredients may even reactwith components present on the skin surface.

Controlling the odour emanating from the body is an important cosmeticbenefit. Indeed, perfume has been used for centuries as a way to enhanceoverall personal appearance. In particular, studies have shown thatattractiveness could be influenced by smell. It is unclear whetherodours alter the visual perception of facial features or, alternatively,how faces are emotionally evaluated by the brain, but what is clear isthat pleasant odour and facial attractiveness integrate into one jointemotional evaluation. Furthermore, to the extent that personalwell-being can be affected by how attractive an individual feels, or howattractive an individual is perceived to be by others, control of bodyodour can foster positive social impact within communities as it canchange the way that human beings perceive one another.

The use of perfume to mask malodour or to create a pleasant odouremanating from the skin is a benefit that is particularly useful in thecontext of cosmetic products, because pleasant odours may help toreinforce the impression of health and attractiveness that theseproducts are designed to impart.

In the design of perfumes, such as fine fragrances, a perfume'sperformance is assessed by the way in which it evaporates from paperblotters. These blotters are cellulosic in nature and provide anevaporation surface that is not representative of the skin. Theexperienced perfumer will understand that the surface of a blotter isdissimilar to the surface of human skin, and will make allowances forthis in perfume design. But the perfumer's task is complicated if themicrobiota is unbalanced and changes unpredictably over time, alteringthe skin's surface chemistry in a similarly unpredictable way.

Overcoming these technical constraints in order to design perfumes thatevaporate more predictably from the surface of the skin is an unmetneed.

Surprisingly, the applicant has discovered that using the cosmeticingredient of the present invention in conjunction with a perfumecomposition, one is able to balance the skin microbiota over a period oftime, and thereby provide a more consistent surface from which a perfumecomposition can emanate in a more predictable manner.

Still further, by employing the cosmetic ingredient of the presentinvention to balance the skin's microbiota, the perfumer is able to makea more considered selection of perfume ingredients in order to achieve aparticular hedonic effect or to alter the temporal profile of a perfumecomposition, for example by enhancing the initial bloom of certainperfume ingredients or rendering other perfume ingredients moresubstantive and long-lasting on the skin's surface.

Accordingly, the invention provides in another of its aspects a method amodulating the evaporation of a perfume composition applied to thesurface of the skin, the method comprising the step of applyingsimultaneously, sequentially or separately the cosmetic ingredient tothe skin.

In particular embodiments of the invention, the method of modulating theevaporation of the perfume composition enhances the initial perceptionof a perfume composition on the skin or enhances retention of a perfumecomposition on the skin, or both.

Nowadays, many people try to avoid certain commonly employedantiperspirant active ingredients, such as aluminium or zirconium saltsor bactericides and bacteriostatic agents, since they are perceived asbeing unfriendly to human skin, or to populations of micro-floracontained on human skin. Consequently, consumers increasingly wish totransition from antiperspirant to more skin friendly deodorant products.However, they are in some cases dissuaded from doing so because of thepossibility of breakthrough malodour in the transition period. Applicantbelieves that such breakthrough malodour may be the result of changes tothe microbiome.

The cosmetic ingredient of the present invention is able to improve theefficacy and long-lastingness of a deodorant composition. In particular,it provides a better malodour reduction and an increased fragranceintensity when used in a deodorant composition (see example 3 below).This is especially advantageous for people changing from anantiperspirant to a deodorant composition, as it reduces the differencebetween the effects of the two types of composition.

Thus, in another aspect, the present invention provides a method forimproving the efficacy and/or long-lastingness of a deodorantcomposition, comprising the step of adding the cosmetic ingredient tothe deodorant composition.

The present invention further relates to a method of facilitating thetransition from an antiperspirant to a deodorant composition byproviding a deodorant composition comprising the cosmetic ingredient ofthe present invention. The present invention further relates to the useof the cosmetic ingredient in an antiperspirant or a deodorantcomposition, and in particular in a deodorant composition comprising noaluminium or zirconium salts.

In another aspect of the invention, there is provided an augmentedperfume composition comprising a perfume composition and the cosmeticingredient defined above.

In a particular embodiment of the invention, the augmented perfumecomposition is a fine fragrance.

In a more particular embodiment of the invention, the augmented perfumecomposition is a fine fragrance comprising a perfume composition, thecosmetic ingredient defined above, and ethanol.

The cosmetic ingredient is useful to condition or balance the skin'smicroflora, and so it is somewhat counter-intuitive to use it in atopical formulation containing large amounts of ethanol, such as a finefragrance formulation, because one might expect the ethanol to inhibitor kill the microflora. However, in a fine fragrance composition, thecosmetic ingredient of the present invention can help off-set any damagecaused to the mircoflora by the ethanol.

As used herein, evaporation of a perfume composition is considered to bemodulated if the headspace of the perfume composition above the surfaceof skin is altered relative to the headspace of the same perfumecomposition by simultaneous, sequential or separate application of thecosmetic ingredient to the skin surface. By analogy, an augmentedperfume composition is a perfume composition that exhibits a modulatedheadspace by virtue of it containing the cosmetic ingredient definedabove.

The discovery that the cosmetic ingredient of the present invention iscapable of balancing the skin's microbiota enables the skilled perfumerto create bespoke perfume compositions that contain perfume ingredientsselected to evaporate at a pre-defined rate having regard to thecompositional make-up of the microbiota and related chemical environmenton the skin's surface.

The cosmetic ingredient of the present invention may be appliedtopically to the skin to provide all manner of beneficial effects,including those related to the preservation of skin beauty over time, aswell as the perfume benefits referred to above. In particular, thecosmetic ingredient may be employed in all manner of cosmetic productsuseful in skin resurfacing; skin renewal activation; skin regeneration;treatment of dry skin; anti-aging; anti-wrinkling; skin moisturizing andhydration; skin barrier reinforcement; natural skin exfoliation; andskin microbiota protection.

Particular cosmetic benefits that can be achieved through theapplication of the cosmetic ingredient of the present invention include:improving skin appearance, improving skin feel, increasing the thicknessof one or more layers of the skin, increasing the elasticity of theskin, increasing the resiliency of the skin, increasing the firmness ofthe skin, reducing an oily appearance of the skin, reducing a shinyappearance of the skin, reducing a dull appearance of the skin,increasing the hydration of the skin, creating a moisturizing effect onthe skin, reducing the appearance of fine lines, reducing the appearanceof wrinkles, improving skin texture, improving skin smoothness,improving skin exfoliation, improving skin desquamation, plumping theskin, improving skin barrier properties, improving skin tone, reducingthe appearance of redness, reducing the appearance of skin blotches,improving the brightness of the skin, improving the radiance of theskin, and improving the translucence of the skin.

The cosmetic ingredient according to the present invention may beformulated in a formulation suitable for topical application to theskin.

The formulation suitable for topical application may be all manner ofpersonal care products of the leave-on or rinse-off variety, including,but not limited to, make-up preparations, fine fragrances, colognes,toilet water, soaps and detergents, deodorants, douches, femininehygiene deodorants, shaving preparations, skin care preparations, e.g.cleansing, depilatories, face and neck, body and hand, moisturizers,suntan preparations, e.g. gels, creams, and liquids, and indoor tanningpreparations.

The formulation suitable for topical application may contain at leastone cosmetically acceptable excipient.

In some embodiments, a cosmetically acceptable excipient refers to acosmetically acceptable material, composition, or vehicle, such as aliquid or solid filler, diluent, solvent, or encapsulating material. Insome embodiments, the excipient is cosmetically acceptable in the senseof being compatible with the other ingredients of a topical formulation,and suitable for use in contact with the skin. Any excipients commonlyused in the preparation of formulations suitable for topical applicationto the human skin, such as cosmetic preparations and fine fragrances,may be employed in the present invention. Suitable excipients include,but are not limited to, ingredients that can influence organolepticproperties, penetration of the skin, and the bioavailability of thecosmetic ingredient. More specifically, they include liquids, such aswater, oils or surfactants, including those of petroleum, animal, plantor synthetic origin, such as and not restricted to, peanut oil, soybeanoil, mineral oil, sesame oil, castor oil, polysorbates, sorbitan esters,ether sulfates, sulfates, betaines, glycosides, maltosides, fattyalcohols, nonoxynols, poloxamers, polyoxyethylenes, polyethyleneglycols, dextrose, glycerol, digitonin, and the like.

The formulation for topical application to the skin may take anyphysical form.

The formulation may be in the form of a liposome composition, mixedliposomes, oleosomes, niosomes, ethosomes, milliparticles,microparticles, nanoparticles and solid-lipid nanoparticles, vesicles,micelles, mixed micelles of surfactants, surfactant-phospholipid mixedmicelles, millispheres, microspheres and nanospheres, lipospheres,millicapsules, micro-capsules and nanocapsules, as well asmicroemulsions and nanoemulsions, which can be added to achieve agreater penetration of the cosmetic ingredient.

The formulation may be produced in any solid, liquid, or semi-solid formuseful for application to the skin topically. Thus, these preparationsof topical application include, but are not restricted to, creams,multiple emulsions, such as and not restricted to, oil and/or siliconein water emulsions, water-in-oil and/or silicone emulsions,water/oil/water or water/silicone/water type emulsions, andoil/water/oil or silicone/water/silicone type emulsions,micro-emulsions, emulsions and/or solutions, liquid crystals, anhydrouscompositions, aqueous dispersions, oils, milks, balsams, foams, aqueousor oily lotions, aqueous or oily gels, cream, hydro-alcoholic solutions,hydro-glycolic solutions, hydrogels, liniments, sera, soaps, face masks,serums, polysaccharide films, ointments, mousses, pomades, pastes,powders, bars, pencils and sprays or aerosols (sprays), includingleave-on and rinse-off formulations.

There now follows a series of examples that serve to further illustratethe invention.

EXAMPLE 1: PREPARATION OF COSMETIC INGREDIENT

Dried Orobanche rapum, collected manually in Lozére, France, was driedfor 1 month in shady conditions before being ground into powder. 1 kg ofOrobanche rapum powder was extracted in 6 kg of 96% ethanol in theabsence of water. Thereafter, the solid material was separated from theliquid extract by filtration over a cellulosic filter (0.7 micron).

The filtrate was combined with an equal mass of 1,3-propanediol beforethe ethanol was removed by vacuum evaporation. The resultant extract in1,3-propanediol was filtered over a charcoal filter. The decolourizedextract was adjusted to 1 wt % dry matter and filtered on a 0.3 micronfilter.

Analysis by 2D NMR identified two major peaks that representing 18.9 wt% (based on total weight of dry matter), which peaks correspond tocrenatoside and acteoside.

EXAMPLE 2: METAGENOMIC ANALYSIS

Clinical assessments of the cosmetic ingredient formed according toExample 1 were carried out on human volunteers.

A double blind and placebo-controlled clinical evaluation was carriedout with 19 women (Age: between 18 and 50 years old, mean age: 39.5years) having dry skin (corneometer value below 40 AU). All of thesubjects participating in the study gave their informed consent signedat the beginning of the study. The measurements were done after 14 and28 days of use with a test formula (set forth below) containing 0.5% ofthe extract obtained in accordance with Example 1. A placebo consistedof the same formulation (below) without the extract.

In this study, the evolution of skin metagenome was followed. The bodyarea studied was the forearm.

The test formulation, in the form of a cream, contained the followingingredients:

WATER, CAPRIC/CAPRYLIC TRIGLYCERIDE, CETEARYL WHEAT STRAW GLYCOSIDES,CETEARYL ALCOHOL, EXTRACT OF EXAMPLE 1, PHENOXYETHANOL, DIMETHICONE,METHYL PARABEN, PROPYL PARABEN, ETHYL PARABEN, FRAGRANCE, HEXYLCINNAMAL, BUTYLPHENYL METHYLPROPIONAL, CITRONELLOL, ALPHA ISOMETHYLIONONE, HYDROXYISOHEXYL 3-CYCLOHEXENE CARBOXALDEHYDE, SODIUM HYDROXIDE.

A placebo formulation was identical to the test formulation (above)except that the extract of Example 1 was omitted.

Skin samples of cutaneous microflora were collected from the forearms ofhealthy volunteers (50 cm²) by a non-invasive swabbing method usingsterile swabs moistened with a sterile solution of 0.15 M NaCl. Swabswere transferred at −20° C. and kept frozen until DNA extraction.

DNA extraction was performed using the PowerLyzer® PowerSoil® DNAIsolation Kit (MO BIO Laboratories, Inc., Carlsbad, USA), with thefollowing modifications. The tip of each swab was detached with asterile surgical blade and transferred to a 1.5 mL tube to which 750 μLof Bead Solution had been added. The sample biomass was re-suspended bystirring and pipetting and the biological suspension was transferred toa bead beating tube. The remaining steps were performed according to themanufacturer instructions.

Sequencing and DNA analysis was carried out as described below.

16S rRNA gene sequencing:

Sequencing was performed with the MiSeq device (Illumina, Inc., SanDiego, Calif., USA) through a 500 cycles paired-end run, targeting theV3V4 16S variable regions using the following primers: 16S-Mi341Fforward primer 5′-CCTACGGGNGGCWGCAG-3′ and 16S-Mi805R reverse primer5′-GACTACHVGGGTATCTAATCC-3′, producing about 460 bp amplicons.

PCR1s were performed as follows: 8 μL of template DNA (0.2 ng) weremixed with 5 μL of each reverse and forward primer (1 μM), 5 μL of KAPAHiFi Fidelity Buffer (5×), 0.8 μL of KAPA dNTP Mix (10 mM each), 0.7 μLof RT-PCR grade water (Ambion), and 0.6 μL of KAPA HiFi hotstart Taq (1U/μL), for a total volume of 25 μL. Each amplification was duplicated,and duplicates were pooled after amplification. PCR1 cycles consisted of95° C. for 3 min and then 32 cycles of 95° C. for 30 s, 59° C. for 30 s,and 72° C. for 30 s, followed by a final extension at 72° C. for 3 min,with a BioRad CFX1000 thermocycler. Negative and positive controls wereincluded in all steps to check for contamination. All duplicate poolswere controlled by gel electrophoresis, and amplicons were quantifiedusing fluorometry.

Libraries ready for analysis were then produced following the Illuminaguidelines for 16S metagenomics libraries preparation. Briefly, the PCR1amplicons were purified and controlled using an Agilent 2100 Bioanalyzer(Agilent Technologies, Santa Clara, USA). To enable the simultaneousanalysis of multiple samples (multiplexing), Nextera® XT indexes(Illumina) were added during PCR2 using between 15 to 30 ng of PCR1amplicons. PCR2 cycles consisted of 94° C. for 1 min and then 12 cyclesof 94° C. for 60 s, 65° C. for 60 s, and 72° C. for 60 s, followed by afinal extension at 72° C. for 10 min. Indexed libraries were purified,quantified and controlled using an Agilent 2100 Bioanalyzer. Validatedindexed libraries were pooled in order to obtain an equimolar mixture.

The run (500 cycles) was achieved on MiSeq sequencer (Illumina) usingthe MiSeq Reagent Kit v3 600 cycles (Illumina). The sequencing runproduced an output of 25 million of paired-end reads of 250 bases, i.e.up to 6 Gigabases. The libraries and the MiSeq run were performed byLibragen, at the GeT-PlaGe platform (INRA, Auzeville, France).

After MiSeq run, raw data sequences were de-multiplexed andquality-checked to remove all reads with ambiguous bases. Indexes andprimer sequences were then trimmed, and the forward and reversesequences were paired. The paired-sequences were then treated usingin-house pipeline to remove chimeras and reads with PCR errors and tosplit sequences into Operational Taxonomic Unit (OTU) at a 1%dissimilarity level. Good quality binned paired-sequences were mapped tothe SILVA SSU Ref database (Release 123; https://www.arb-silva.de/) fortaxonomic assignation. Data were then normalized, and compared usingWhite's non-parametric test (White et al., 2009).

The results are shown in the following two tables.

TABLE 1 Forearm microbiota phyla composition after 14 days of treatmentwith Vehicle and Vehicle + Active Vehicle Vehicle + Active D0-Vehicle:D14-Vehicle: D0-Active: D14-Active: mean rel. mean rel. mean rel. meanrel. Phylum frequ. (%) frequ. (%) p-values frequ. (%) frequ. (%)p-values Actinobacteria 38.71 34.42 0.387 38.05 33.88 0.443Proteobacteria 26.78 22.27 0.372 27.25 25.73 0.775 Firmicutes 26.1937.00 0.025 26.10 29.47 0.486 Bacteroidetes 6.18 3.61 0.144 6.62 3.940.176 Cyanobacteria 1.15 1.91 0.496 1.36 6.04 0.272 Saccharibacteria0.14 0.09 0.092 0.09 0.08 0.852 Chloroflexi 0.11 0.09 0.794 0.07 0.140.061

Table 1 lists the main phyla (in terms of relative abundance) found inthe study. Table 1 also demonstrates that the microbiota evolvesdifferently depending on whether skin was treated with the testformulation (“Vehicle+Active”) or the placebo (“Vehicle”) formulation.At the phylum level, skin treated with the placebo formulation exhibiteda significant increase of Firmicutes between D0 and D14, whereas thelevels of other phyla remained substantially stable. On the other hand,for skin treated with the test formulation, levels of all phyla,including Firmicutes, remained substantially stable.

It can be concluded that the cosmetic ingredient stabilises themicrobiota and maintains the essential balance of the skin microflora.

TABLE 2 Comparison of the relative abundances of Finegoldia magna at D0and D14 Vehicle + Active 0.5% Vehicle D0-Active: D14-Active: D0-Vehicle:D14-Vehicle: mean rel. mean rel. mean rel. mean rel. Genus frequ. (%)frequ. (%) p-value frequ. (%) frequ. (%) p-value Finegoldia 3.1 1.30.043 1.9 2.1 0.741

Table 2 shows the levels of Finegoldia magna on skin treated with testformulation and placebo (Vehicle) over a 14-day period.

Finegoldia magna is a species that is a normal inhabitant of human skinand is most frequently Gram positive cocci isolated from infectedlesions. F. magna is implicated in mono- and poly-microbial infectionsof skin, bone, heart and meninges. A case of toxic shock syndrome causedby F. magna has also been reported in the literature.

Results of the study show that skin treated with the test formulation(“Active 0.5%”) exhibits a significant reduction in the relativeabundance of Finegoldia over the 14 day test period (−58%). On the otherhand, when skin is treated with the placebo formulation (“Vehicle”), thelevel of Finegoldia remains stable. These finding suggest that thecosmetic ingredient inhibits the growth of opportunistic pathogens.

EXAMPLE 3: DEODORANT COMPOSITION

In order to assess the effect of the cosmetic ingredient of the presentinvention on deodorant efficacy, a double blind study on 10 volunteerswas conducted. All volunteers were antiperspirant users.

The volunteers had to stop using their usual antiperspirant. Instead,they applied a first test deodorant composition on their left underarmand a second test deodorant composition on their right underarm eachmorning.

At the end of a five-day test period, the volunteers made aself-assessment of the malodour and the fragrance for each side.

Both test deodorant compositions were provided in the form of a sprayand contained the following standard deodorant base:

1 wt % Octyl Dodecanol 36 wt %  Ethyl alcohol 96% 2 wt % PropyleneGlycol 1 wt % Perfume 60 wt %  Propellant (Isobutane, Propane)

The above ingredients were mixed without the propellant and supplementedwith 1.75 wt % of a fragrance composition. For one of the two testdeodorant compositions, 0.5 wt % of the cosmetic ingredient of thepresent invention was added. The mixtures were filled into a spraycontainer and charged with the propellant.

The self-assessment showed that 9 out of the 10 volunteers found thatthe test deodorant composition containing the cosmetic ingredient of thepresent invention provided a better malodour reduction and an increasedfragrance intensity.

Thus, the cosmetic ingredient of the present invention is able toimprove the long-lastingness of a deodorant composition.

1. A cosmetic ingredient comprising crenatoside and acteoside.
 2. Thecosmetic ingredient according to claim 1 in the form of a plant extractcomprising crenatoside and acteoside.
 3. The cosmetic ingredientaccording to claim 2, comprising the extract and a hydric organicsolvent.
 4. The cosmetic ingredient according to claim 3, wherein thehydric organic solvent includes glycerol, a glycol or 1,3-propanediol.5. A formulation adapted for topical application to the skin comprisinga cosmetic ingredient according to claim
 1. 6. The formulation accordingto claim 5, comprising a perfume composition containing at least oneperfume ingredient.
 7. The formulation according to claim 5, selectedfrom the group consisting of: a cosmetic cream, cosmetic lotion,cosmetic body spray, cosmetic serum, a deodorant or antiperspirant, or afine fragrance.
 8. A method of modulating the evaporation of a perfumecomposition applied to the skin, the method comprising the step of:applying simultaneously, sequentially or separately the cosmeticingredient according to claim 1 to the skin.
 9. A method of improvingthe efficacy and/or long-lastingness of a deodorant composition,comprising the step of adding the cosmetic ingredient according to claim1 to the deodorant composition.
 10. A method of facilitating thetransition from an antiperspirant to a deodorant composition byproviding a deodorant composition comprising the cosmetic ingredientaccording to claim
 1. 11. A deodorant composition or an antiperspirantcomposition which comprises the cosmetic ingredient of claim 1, andfurther wherein the deodorant composition or antiperspirant compositioncomprises no aluminium salts or zirconium salts.